Knockout of Bptf in MEFs was accomplished by infecting litter mate wild type and conditional Bptf (-/Floxed genotype) MEF lines with an adenovirus expressing CMV-Cre. Infection was performed in 6 well plates at sub-confluence (2×105 cells) with 1 ml growth media containing 45 µl Adenovirus @ 1×1010 PFU/ml. Cells were grown for two days in the presence of virus. Following infection cells were divided into 5 wells of a 6 well plate. Cell numbers were recorded every 24 hours with changes in media occurring every 48 hours. A sample of cells was removed at day 4 for Western analysis of Bptf protein levels. Experiment was repeated with two wild type and conditional knockout MEF lines from littermate embryos.
